LOCUS       sequence-297161-        2250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TwinStrep-GFP11 C-terminal Tag Donor Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2250)
  AUTHORS   Pullmann P, Knorrscheidt A, Munch J, Palme PR, Hoehenwarter W, 
            Marillonnet S, Alcalde M, Westermann B, Weissenborn MJ
  TITLE     A modular two yeast species secretion system for the production and 
            preparative application of unspecific peroxygenases.
  JOURNAL   Commun Biol. 2021 May 12;4(1):562. doi: 10.1038/s42003-021-02076-3.
  PUBMED    33980981
REFERENCE   2  (bases 1 to 2250)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2021-05-12"; volume: "4(1):562. doi"; pages: " 
            10.1038/s42003-021-02076-3"
COMMENT     Sequence Label: pAGM9121_TwinStrep-GFP11
FEATURES             Location/Qualifiers
     source          1..2250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             285..1076
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1417..2005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1906..1925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2064..2069,2070..2093,2094..2129,2130..2153)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2184..2231
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
ORIGIN
        1 gtgccgaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
       61 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
      121 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
      181 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      241 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg gaagcggtga
      301 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
      361 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
      421 gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
      481 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
      541 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
      601 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
      661 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
      721 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
      781 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
      841 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
      901 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
      961 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1021 tggaagaatt tgtccattac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc
     1081 tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga
     1141 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt
     1201 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgctgt cagaccaagt
     1261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgca
     2041 ctctgtggtc tcattcgggt ggttctgctt ggtcacatcc acaatttgaa aaaggtggag
     2101 gttcaggtgg aggttcgggt ggttctgctt ggtcacatcc acaatttgaa aaagatggtg
     2161 gttctggtgg tggttctact agtcgtgatc atatggttct tcatgaatat gttaatgctg
     2221 ctggtattac ttgagcttcg agaccacgaa
//
